Bigarré L, Baud M, Cabon J, Antychowicz J, Bergmann S M, Engelsma M, Pozet F, Reichert M, Castric J
Pathologie Virale des Poissons, AFSSA, Technopôle Brest-Iroise, Plouzané, France.
J Virol Methods. 2009 Jun;158(1-2):51-7. doi: 10.1016/j.jviromet.2009.01.023. Epub 2009 Feb 4.
To date, all the isolates of Cyprinid herpesvirus type-3 (CyHV3) responsible for serious outbreaks in carps Cyprinus carpio have been found to be very similar or identical on the basis of DNA sequences of a few reference genes. However, two genetic lineages (U/I and J) are distinguished by full-length genome sequencing. Two molecular markers presenting genetic variations were targeted for developing a duplex PCR assay able to distinguish CyHV3-U/I from CyHV3-J while avoiding DNA sequencing. The method was validated on a series of 42 samples of infected carps from France, The Netherlands and Poland collected from 2001 to 2008. Among these samples, both the U/I and J genotypes were identified, but also a third genotype representing a genetic intermediate between U/I and J for one of the two molecular markers. A classification of CyHV3 genotypes, based on the alleles of the two molecular markers, is proposed. The assay is easy to perform and provides a genotype information with samples moderately or highly concentrated. This tool should improve our knowledge regarding the present distribution and future diversification of this emerging virus.
迄今为止，根据少数参考基因的DNA序列，所有导致鲤鱼（Cyprinus carpio）严重疫情爆发的鲤疱疹病毒3型（CyHV3）分离株都非常相似或完全相同。然而，通过全长基因组测序可区分出两个遗传谱系（U/I和J）。针对两个呈现遗传变异的分子标记开发了一种双重PCR检测方法，该方法能够区分CyHV3-U/I和CyHV3-J，同时避免进行DNA测序。该方法在2001年至2008年从法国、荷兰和波兰采集的42份感染鲤鱼样本系列上进行了验证。在这些样本中，不仅鉴定出了U/I和J基因型，还鉴定出了第三种基因型，该基因型在两个分子标记之一上代表U/I和J之间的遗传中间类型。基于两个分子标记的等位基因，提出了CyHV3基因型的分类方法。该检测方法易于操作，并且能为中度或高度浓缩的样本提供基因型信息。这个工具应该能增进我们对这种新出现病毒当前分布和未来多样性的了解。
